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The Type III Secretion System-Related CPn0809 from Chlamydia pneumoniae

机译:肺炎衣原体与III型分泌系统有关的CPn0809

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Chlamydia pneumoniae is an intracellular Gram-negative bacterium that possesses a type III secretion system (T3SS), which enables the pathogen to deliver, in a single step, effector proteins for modulation of host-cell functions into the human host cell cytosol to establish a unique intracellular niche for replication. The translocon proteins located at the top of the T3SS needle filament are essential for its function, as they form pores in the host-cell membrane. Interestingly, unlike other Gram-negative bacteria, C. pneumoniae has two putative translocon operons, named LcrH_1 and LcrH_2. However, little is known about chlamydial translocon proteins. In this study, we analyzed CPn0809, one of the putative hydrophobic translocators encoded by the LcrH_1 operon, and identified an ‘SseC-like family’ domain characteristic of T3S translocators. Using bright-field and confocal microscopy, we found that CPn0809 is associated with EBs during early and very late phases of a C. pneumoniae infection. Furthermore, CPn0809 forms oligomers, and interacts with the T3SS chaperone LcrH_1, via its N-terminal segment. Moreover, expression of full-length CPn0809 in the heterologous host Escherichia coli causes a grave cytotoxic effect that leads to cell death. Taken together, our data indicate that CPn0809 likely represents one of the translocon proteins of the C. pneumoniae T3SS, and possibly plays a role in the translocation of effector proteins in the early stages of infection.
机译:肺炎衣原体是一种细胞内革兰氏阴性细菌,具有III型分泌系统(T3SS),可使病原体在单个步骤中将调节宿主细胞功能的效应蛋白转运到人宿主细胞胞浆中,从而建立一种独特的细胞内复制位。位于T3SS针状细丝顶部的translocon蛋白对其功能至关重要,因为它们在宿主细胞膜中形成孔。有趣的是,与其他革兰氏阴性细菌不同,肺炎衣原体具有两个推定的translocon操纵子,命名为LcrH_1和LcrH_2。然而,关于衣原体转运蛋白的了解甚少。在这项研究中,我们分析了由LcrH_1操纵子编码的疏水性转运子之一CPn0809,并确定了T3S转运子的“ SseC样家族”结构域特征。使用明场和共聚焦显微镜,我们发现CPn0809在肺炎衣原体感染的早期和晚期都与EB有关。此外,CPn0809形成低聚物,并通过其N末端片段与T3SS分子伴侣LcrH_1相互作用。而且,全长CPn0809在异源宿主大肠杆菌中的表达引起严重的细胞毒性作用,导致细胞死亡。两者合计,我们的数据表明CPn0809可能代表肺炎衣原体T3SS的一种translocon蛋白,并且可能在感染的早期阶段就参与了效应蛋白的转运。

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